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AB C D
E F
POTENTIAL BENEFITS OF DOCOSAHEXAENOIC ACID IN A MOUSE MODEL OF LUNG CANCER-RELATED CACHEXIA
IC007
Raquel D.S. Freitas 1,4, Kesiane M. da Costa 1, Fernanda L. Rost 3,4, Maria Eduarda A. Amaral 2,3, Maria M. Campos 1,2,3,4
1 Programa de Pós-graduação em Medicina e Ciências da Saúde - Escola de Medicina - PUCRS; 2 Programa de Pós-graduação em Biologia Celular e Molecular - Escola de Ciências - PUCRS; 3 Curso de Graduação em Odontologia – Escola de Ciências da Saúde – PUCRS; 4 Centro de Toxicologia e Farmacologia – Escola de Ciências da Saúde – PUCRS.
Cancer cachexia (CC) is a multifactorial syndrome, characterized by muscle atrophy, loss of adipose tissue and reduced quality of life that cannot be fully reversed by nutritional therapy (Lancet Oncol. 12:489. 2011).
About 50% of cancer deaths are related to tumors associated with
cachexia (Nat Rev Dis Primers. 4:1-18. 2018). N-3 fatty acid eicosapentaenoic acid (EPA) supplementation is widely
used for management of CC (Br J cancer. 105:1469. 2011; Cancer Metastasis Rev. 34:359. 2015), although, other n-3 fatty acids, such as docosahexaenoic acid (DHA), have not been fully investigated.
AIM
This study assessed the effects of DHA treatment in a mouse model of cancer cachexia.
METHODS
in v
itro
C57/BL6 male mice (20 – 25g/8 weeks ) CEUA/PUCRS approval number: 15/7164
Euthanasia Removal of tissues for
weight and histological analysis
Blood collection for hematological analysis
Lewis lung carcinoma cell line LL/2 (LLC1) was used.
Cells were cultivated in DMEM 10% SFB, at 37oC, 5% CO2.
Number of cells was counted using a Neubauer chamber.
Cells were centrifuged for 10 min at 3000 RPM.
Cells were resuspended in 100 µL of PBS.
5x106 cells were injected in the dorsal region of each mouse.
0 7 21
Cachexia model
induction
Behavioral tasks
• Control
• DHA 1 µmol/kg i.p.
• LLC
• LLC + DHA 1 µmol/kg i.p.
EXPERIMENTAL GROUPS
in v
ivo
Behavioral tasks
DHA i.p. treatment
RESULTS
Fig 1. Evaluation of exploratory activity shown as distance (A) ambulatory movement, (B) rearing, (C) speed, (D) traveled distance, after 21 days of LLC-induced cachexia and 15 days of systemic DHA treatment. Effects of DHA treatment on (E) grip strength analysis, and (F) motor coordination tested by rota-rod apparatus after LLC-cachexia induction. Each column represents the mean ± SEM of 8-10 mice per experimental group. ##P<0.01 significantly different from control group;
RESULTS
Fig 2. Effects of parenteral administration of DHA in healthy mice and in LLC-cachectic animals on body weight and glycemia levels. (A) Analysis of body weight variation (tumor-free carcass weight minus initial weight prior to tumor induction) throughout 21 days of cachexia development, (B) Tumor-free carcass weight and (C) Glycemia levels after 6 h-fasting period. Each column represents the mean ± SEM of 6-12 mice per experimental group. #P<0.05 significantly different from LLC-cachectic group; *P<0.05 significantly different from control group.
Fig 3. Effects of DHA i.p. treatment on epididimal fat (A) retroperitoneal fat, (B) intrascapular fat (C) inguinal fat, (D) weights of LLC-cachectic and healthy mice. Each column represents the mean ± SEM of 4-12 mice per experimental group. *P<0.05 significantly different from control group.
Fig 5. Effects of LLC-cachexia on (A) hematocrit levels, (B) neutrophils-to-lymphocytes ratio, and (C) spleen weight. Each column represents the mean ± SEM of 6-12 mice per experimental group. *P<0.05 significantly different from control group.
INTRODUCTION
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Fig 6. Evaluation of (A) retroperitoneal fat and (B) intrascapular fat adipocyte área, and (C) frequency distribution of isWAT adipocyte area. Each column represents the mean ± SEM of 3-8 mice per experimental group. *P<0.05 significantly different from control group. Representative images of intrascapular fat: (A) Control, (B) DHA 1 µmol/kg i.p., (C) LLC and (D) LLC + DHA 1µmol/kg i.p.
CONCLUSION
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Fig 4. Effects of DHA i.p. treatment on (A) gastrocnemius, (B) soleus, and (C) tibialis anterior weights of LLC-cachectic and healthy mice. Each column represents the mean ± SEM of 8-12 mice per experimental group. *P<0.05 significantly different from control group.
A
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Parenteral administration of DHA showed some beneficial effects on the mouse model of lung cancer cachexia, specifically regarding adipose tissue loss. Additional analysis are in progress in order to identify the mechanisms of DHA in this model of cancer-related cachexia.
FINANCIAL SUPPORT FINEP-PUCRSINFRA #01.11.0014-00